Geographical Distribution of Trypanosoma cruzi Genotypes in Venezuela

<p>Chagas disease is caused by a protozoan parasite called<br>Trypanosoma cruzi. T. cruzi infects a wide variety of<br>mammal species in Latin America as well as man, and is<br>spread by multiple species of blood sucking triatomine<br>insect vectors. The presence of genetic diversity in T. cruzi<br>in the Americas is well established, with six different major<br>genetic types in circulation. The genetic diversity of T. cruzi<br>in Venezuela is relatively poorly understood. In this work<br>we present the results from the genotyping of over seven<br>hundred isolates from 17 of the 24 states. Our dataset<br>comprises strains isolated from wild and domestic animals,<br>several species of triatomine vector, as well as from human<br>Chagas disease cases, including those associated with oral<br>transmission of T. cruzi. Amongst other findings, our data<br>reveal a surprisingly high frequency of atypical genotypes<br>in humans, particularly TcIV, which has rarely been<br>reported. We evaluate our findings in the context of T.<br>cruzi diversity elsewhere in the Americas, and assess the<br>impact they have on the future of Chagas disease control<br>in Venezuela.</p

Bat species captured in Guarapari municipality, ES state, Brazil.

<p>Bat species captured in Guarapari municipality, ES state, Brazil.</p

Phylogenetic placement of kinetoplastid OTUs detected in bats of Guarapari municipality, ES state, Brazil.

<p>Tree construction from 18S rRNA followed the maximum likelihood (ML) method under Kimura’s two-parameter model and gamma-distributed variation among sites (K2 + G). Numbers at nodes indicate support from 1000 bootstrap replicates. The 14 OTUs clustered into the <i>T</i>. <i>cruzi</i> clade (OTUs 1, 2, 3, 5, 6, 7, 8, 10, 11, 12, 13 and 14), a reptile-associated region (OTU 4) and the <i>B</i>. <i>saltans</i> outgroup (OTU 9).</p

Heatmap of kinetoplastid OTU distribution among bats captured in Guarapari municipality, ES state, Brazil.

<p>Each column represents the infection profile of one infected bat individual. Cell colour denotes the sequence read intensity attributed to each OTU (left), increasing from purple (zero reads) through yellow into red. Bat species and sample IDs are given above/below. Asterisks indicate samples subjected to nested PCR, of which ten also underwent standard PCR (dashed lines). Phylogenetic relationships inferred from 18S rRNA by maximum likelihood (ML) tree construction are plotted at right.</p

Phylogenetic placement of OTU 9 with <i>Bodo saltans</i> among a wider set of trypanosomatid genera.

<p>Tree construction from 18S rRNA followed the maximum likelihood (ML) method under Kimura’s two-parameter model and gamma-distributed variation among sites (K2 + G). Numbers at nodes indicate support from 1000 bootstrap replicates.</p

raw_data_salmon_microbiome

Salmon salar gut microbiome sequences. Samples are identified by their "FLD0001.****_R1.fastq" code in the samples' name

Microsatellite neighbour-joining phylogeny

Unrooted Neighbour-Joining tree based on DAS values between MLGs generated from 199 sylvatic Bolivian TcI clones

Mitochondrial maximum-likelihood phylogeny

Maximum-likelihood tree constructed from concatenated maxicircle sequences for 78 sylvatic Bolivian TcI clones and 24 additional TcI isolates from across the Americas

Table S2: Panel of microsatellite loci and primers

Panel of microsatellite loci and primers employed in this study